Method for Preparing Nanolipids with Encapsulated Alcohol

ABSTRACT

A method for preparing ethanol-containing nanolipid particles, which can be used in food products, frozen desserts, or beverages. The method comprises nanolipidic vehicles in which ethanol-containing substances are encapsulated, said ethanol-containing nanolipidic vehicles can be combined with food products, desserts or beverage ingredients including those that are subsequently frozen. The food product, dessert or beverage can remain in a frozen state during consumption by an individual. A composition comprising ethanol-containing nanolipid particles, which can be used in food products, frozen desserts, or beverages.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.13/840,810 filed Mar. 15, 2013, which is a continuation-in-part of U.S.application Ser. No. 12/772,838 filed May 3, 2010, which is acontinuation of U.S. application Ser. No. 11/644,281 filed Dec. 22,2006, and which claims the benefit of U.S. Provisional Application No.60/755,171 filed Dec. 30, 2005.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

Not applicable.

TECHNICAL FIELD OF INVENTION

This invention relates to the field of encapsulation of ethanol innanolipidic particles.

BACKGROUND OF THE INVENTION

Frozen foods, particularly frozen desserts and frozen beverages, arevery popular with consumers. Frozen desserts, such as ice creams andsorbets are consumer favorites, and are frequently flavored withliqueurs such as Grand Marnier® and Kahlua®. Frozen beverages, such asmargaritas and piña coladas, are also popular. Attempts to provide suchfrozen desserts and beverages with an ethanol content comparable to thenon-frozen counterparts has been met with limited success due to thesubstantially lower freezing point of ethanol as compared to water-basedproducts.

The freezing point of pure water is 0° C. (32° F.). The freezing pointof pure ethanol is −114° C. (−173.2° F.). The freezing point of ethanolcontaining products will fall into the range between these two limits,with the freezing point of an alcohol-containing food product dependingupon the percentage of alcohol (ethanol) in the final product. Practicaland physical limitations prevent the use of commercial freezingmechanisms capable of maintaining high-ethanol content foods attemperatures low enough to stay frozen. Most freezing apparatuses have afunctional range for freezing a food product, and consumer safety willalso dictate a temperature range wherein frozen foods may be safelyingested. Freezing food products with alcohol ranging up to 15% in thefinal concentration requires freezing at temperatures substantiallybelow the freezing point of water.

This decreased freezing point has long been understood to a limitingfactor in the ability to make products containing ethanol which canremain frozen long enough for an individual to reasonably consume theproduct while it remains in the frozen state. Various means have beenemployed to overcome this obstacle, most of which have involved theaddition of stabilizing materials, such as gels or agar, to the foodproduct. Even then, there has been limited success.

Incorporating passenger molecules, such as pharmaceutical activeingredients, in lipid vesicles such as liposomes has been reported inthe prior art. An amphipathic carrier structure denoted as a SolventDilution Microcarrier (“SDMC”) was disclosed in U.S. Pat. No. 5,269,979.In general, the '979 patent described making a plurality of SDMCvehicles by solubilizing an amphipathic material and a passengermolecule in a first quantity of a non-aqueous solvent. Following this, afirst quantity of water was added, forming a turbid suspension. In asubsequent step, a second quantity of non-aqueous solvent was added toform an optically clear solution. The final step of a preferredembodiment was to organize the optically clear solution into SDMCvehicles by mixing with air or a second quantity of water.

In U.S. Pat. No. 5,879,703, a method for preparing a shelf-stableprecursor solution useful for remote encapsulation of active ingredientswas described. In '703, the precursor solution was made by solubilizingan amphipathic material in a non-aqueous solvent. A quantity of waterwas added to the first mixture to form a precursor solutioncharacterized by optical clarity and being monophasic at roomtemperature. The precursor solution could be stored for an extendedperiod of time—and the desired active ingredient added at a later time,perhaps at a remote location, to form a loaded precursor solution. SDMCscould be formed, in preferred embodiments, from the loaded precursorsolution by diluting with water or mixing with air. SDMCs ranged fromabout 230 to about 412 nanometers in size.

Although SDMCs and the shelf-stable precursor solution provided formaking vehicles suitable for delivering active ingredients in a varietyof applications, a need remained for improved vehicles for delivery ofpassenger molecules.

It has now been found that the shelf-stable precursor solution such asdescribed in the '703 patent can be used as a starting material in anovel method which results in vehicles of a smaller size than previouslyreported. The starting material is manipulated by dilution with anon-aqueous solvent, either before or after loading with a passengermolecule, to provide one or more defined populations of nanolipidicparticles (“NLPs”) which range in size from about 1 nanometer to about20 nanometers.

NLP assemblies are formed from the NLPs which range in size from about30 nanometers to about 200 nanometers. In addition, it has been foundthat NLPs can be used in a method for making carrier vehiclepreparations which are mixed smaller and larger carrier vehicles, orhaving a larger mean size of about 200-300 nanometers, but improvedencapsulation of passenger molecules.

SUMMARY OF THE INVENTION

A means has now been found by which ethanol can be effectively,efficiently and economically encapsulated in a nanolipid particle forpossible consumer consumption, such as encapsulation of ethanolmaintained at percentages not previously possible for consumeringestion.

A method for preparing ethanol-containing food products, frozen dessertsand beverages is disclosed using alcohol encapsulated in nanolipidparticles and assemblies. The method comprises nanolipidic vehicles inwhich ethanol-containing substances are encapsulated, saidethanol-containing nanolipidic vehicles are combined with dessert orbeverage ingredients which can subsequently consumed or incorporatedinto food products, such as frozen foods, desserts or beverages. Thesefood items can remain in a frozen state during consumption by anindividual without losing the characteristics of the alcoholencapsulated in nanolipid particles and assemblies.

The method of the claimed invention provides for the encapsulation ofvarious ethanol-containing substances in lipid-based vesicles, saidvesicles being preferably soy-based, which may be added to ingredientsappropriate for consumption in a food product, such as a dessert orbeverage, and the combination may then be frozen by established meansavailable in food service to produce an ethanol-containing food productor frozen food product capable of maintaining a frozen state atconsumer-safe temperatures for a period of time sufficient forconsumption of said product. Additional stabilizing materials do notneed be added to the food product to achieve this result.

DETAILED DESCRIPTION Preparation of Nanolipidic Particles (NLPs) and NLPAssembly Populations

Nanolipidic particles (NLPs) are prepared according to the techniquesset forth in United States Patent Application Publication No.2007/0154539 A1, published Jul. 7, 2007, United States PatentApplication Publication No. 2010/0239686 A1, published Sep. 23, 2010,and United States Patent Application Publication. No. 2012/00195940 A1,published Aug. 2, 2012, which are both herein incorporated by reference.NLPs are prepared from a Shelf-Stable Precursor Stock, preparedaccording to U.S. Pat. No. 5,879,703 which is also incorporated byreference as if fully set forth herein.

NLPs are made from a precursor solution as described in U.S. Pat. No.5,879,703,. As stated in the '703 patent, a precursor solution may bemade by solubilizing an amphipathic material in a first quantity of anon-aqueous solvent appropriate to solubilize the amphipathic materialto form a first mixture. The amphipathic material preferably comprisesphospholipids (PL). Preferred phospholipids comprise one or more of thefollowing phosphatides: phosphatidylcholine (PC),phosphatidylethanolamine (PE), phosphatidic acid (PA) andphosphatidylinositol (PI). In a preferred embodiment, PC, PE, PA and PIare combined. A preferred ratio of PLs useful in the invention isPC:PE:PA:PI of 6.5:2.5:0.7:0.3 in ethanol. Preferably, one gram of PL issolubilized in 5.0-7.5 mL of ethanol solvent.

After dissolution of the amphipathic material, a quantity of water isadded to form a turbid suspension. The amount of water to add isapproximately 9 kg of water to 31 kg of dissolved amphipathic material,but the amount of water can be varied to result in the desired turbidsuspension. A second quantity of non-aqueous solvent, such as ethanol,is added until the turbid suspension is monophasic and has opticalclarity at room temperature. This resulting product is a precursorsolution which is shelf-stable over time.

In the '703 patent, it was disclosed that a precursor solution madeaccording to the process disclosed therein was shelf stable at least upto two years, and perhaps longer, as long as it remains in a monophasiccondition. It has been recently determined that precursor solutions madeby this method are stable for at least eight years, independent ofmanufacturing, location, season, year and lot.

It has now been found that a precursor solution such as disclosed in'703 can be used as a starting material to make nanolipidic particles(NLPs) and NLP assemblies. In '703, the precursor solution was disclosedas being useful for making SDMCs (Solvent Dilution Microcarriers) at alater point in time and, perhaps, a remote location. SDMCs have adiameter of from about 230 to about 412 nm. In contrast, NLPs have amean diameter of from about 1 nm to about 20 nm and NLP assemblies havea mean diameter from about 30 nm to about 200 nm.

Various populations of NLP assemblies may be made for variousapplications. Preferred populations range from about 40-60 nm; about60-80 nm; about 80-110 nm; about 110-140 nm, and about 150-200 nm. NLPassembly populations are generally 20-30% smaller in diameter than SDMCsfor the same passenger molecule.

A slightly larger population or mixed population of carrier vehicles isreferred to herein as ECVs or encapsulating carrier vehicles. Althoughoverlapping the mean diameter of SDMCs, the ECV is made using adifferent method employing NLPs and the result is a carrier vehiclepopulation which has been found to exhibit a higher encapsulatingefficiency. The ECVs are described as having a mean diameter from about200 nm to 300 nm.

To make carriers for passenger molecules, such as NLP populations, NLPassemblies, or ECVs according to the method disclosed in United StatesPatent Application Publication Nos. 2007/0154539, 2010/0239686 and2012/0195940, the precursor solution as previously described in the '703patent is diluted with a suitable solvent or mixed solvent system whichis compatible with the solvent system used in the precursor solution.The concentration of solvent in the NLPs is from about 0.5% to about 14%by volume. This dilution is performed either before or after addition ofthe passenger molecule as will be further described in detail below.

For the present invention, NLPs are prepared by first mixing an aliquotof the aforementioned shelf-stable precursor stock solution with varyingamounts of ethanol to produce NLPs having specific size ranges. TheseNLPs are then diluted with distilled water to achieve the final ethanolconcentration for the claimed composition. Further, these NLPs can beused to form NLP assemblies and carrier vehicle populations utilizingmixed NLP sizes and increased encapsulation of passenger molecules.

The following calculation is used to provide data for the table below:

Neat or Control: (85.0% ethanol in precursor stock)×(1 ml ofpreparation)/20 ml of distilled water=4.3% ethanol

${\frac{85.0\% \mspace{14mu} {EtOH}}{100\mspace{14mu} {mL}\mspace{14mu} {precursor}{\mspace{11mu} \;}{stock}} \times \frac{1\mspace{14mu} {mL}\mspace{14mu} {precursor}\mspace{14mu} {stock}}{20\mspace{14mu} {mL}\mspace{14mu} {total}{\mspace{11mu} \;}{volume}}} = {\% \mspace{14mu} {EtOH}\mspace{14mu} {in}\mspace{14mu} {NLP}\mspace{14mu} {preparation}}$

The resulting sizes and relative ethanol concentrations of the finishedpreparations are tabulated below.

NLPs sizes of 1-20 nm and 10-12 nm are made according to paragraph[0007] of the specification United States Patent Application PublicationNo. 2007/0154539 by dilution of the precursor solution with anon-aqueous solvent.

In Example 1, Table 1 of United States Patent Application PublicationNo. 2007/0154539, the following precursor/ethanol ratios yield NLPassembly populations. The resulting ethanol concentration of the finalproduct has been added to the Table provided in the specification atParagraph [0035] to demonstrate the much lower concentration of ethanolin the NLP assembly populations. Resulting ethanol concentrations weredetermined by adding the ethanol concentration for 1 mL of precursorstock (0.85 mL EtOH/1 mL solution) to the additional amount of ethanolper the chart below and determining the amount of EtOH per mL of theresulting solution, then dividing by the 20 mL water added to make thefinal solution. [For example—0.85 mL EtOH/1 mL stock+2.0 mL EtOHadded=2.85 mL EtOH/3mL total solution→0.95 mL EtOH/1 mL×1 mL/20 mLdiH₂O×100=4.75% final EtOH concentration for a 110 nm size NLPpopulation]

Stock Resulting Precursor Ethanol Ethanol Solution added No PassengerSize Concentration 1 mL   0 mL [Control} 278 nm  4.3% 1 mL  0.2 mL[Mixed population] 242 nm 4.35% 1 mL  0.5 mL [NLP Assembly] 186 nm  4.4%1 mL  0.8 mL [NLP Assembly] 160 nm  4.5% 1 mL  1.0 mL [NLP Assembly] 138nm  4.6% 1 mL  2.0 mL [NLP Assembly] 110 nm 4.75% 1 mL  3.0 mL [NLPAssembly]  98 nm  4.8% 1 mL  5.0 mL [NLP Assembly]  61 nm 4.85% 1 mL10.0 mL [NLP Assembly]  34 nm  4.9%

As evidenced by this example, the ethanol concentration is less than 5%for each of the preparations. Similar ethanol concentrations are alsoproduced with the preparations in Examples 2, 3, 4, 5, 6 & 8. Theremaining examples from the specification have slightly differentethanol concentrations as reported in the sections below.

In Example 7 of United States Patent Application Publication No.2007/0154539, the taste of NaCl is masked by encapsulation in an NLPpreparation. The final ethanol concentration in the NLPs for thisexample is less than 1%. For this preparation, 1 mL of NLP stocksolution is diluted into 100 mL distilled water with a resultingconcentration of 0.9% [(85 mL EtOH/100 mL NLP stock×1 mL/101 mL totalsolution)×100=0.9%]

In Example 9 of United States Patent Application Publication No.2007/0154539, caffeine is the passenger molecule for an NLP preparationwith an ethanol concentration of 14%. For this preparation, 20 mL of NLPstock solution is diluted in 100 mL of distilled water with a resultingethanol concentration of 14%. [(85 mL EtOH/100 mL NLP stock×20 mL/120 mLtotal solution)×100=14%]

In Example 10 of United States Patent Application Publication No.2007/0154539, production of a preloaded NLP preparation containingcaffeine is described. For this preparation, 30 mL of NLP stock solutionis diluted in 50 mL of distilled water with a resulting ethanolconcentration of 0.5%. [(85 mL EtOH/100 mL NLP stock×30 mL/80 mL totalsolution)/62.5×100 =0.5%]

Example 11 of United States Patent Application Publication No.2007/0154539 describes the production of preloaded NLP preparationscontaining lipid soluble vitamins. For this preparation, 1 mL of NLPstock solution is diluted in 50 mL of distilled water with a resultingethanol concentration of 1.7%. [85 mL EtOH/100 mL NLP stock×1 mL/51 mLtotal solution)×100=1.7%]

The range of ethanol concentrations for the NLPs of the invention ofUnited States Patent Application Publication No. 2007/0154539 range froma low of about 0.5% as seen in Example 10 to a high of about 14% as seenin Example 9.

The solvent is selected for biocompatibility if the end use of thecarriers will require that characteristic. The solvent or mixed solventsystem used for dilution must be miscible with the solvents in theprecursor solution and should be effective to disperse rather thandissolve the carriers. Most preferably, the solvent used for dilution isethanol, since it possesses the desired qualities. Ethanol is thesolvent of choice for any end use wherein the particles are foringestion. The dilution is preferably conducted in a sequential orserial manner. For example, a first dilution of 1:10 provides apopulation of carriers, and further serial dilution to about 1:0.5provides a series of populations of carriers.

The size of the carriers in each dilution can be determined by laserlight scattering. Mixed populations of NLPs and larger vesicles may becreated at lower dilutions with the non-aqueous solvent. An appropriateinstrument for this purpose is the Zetasizer 1000 manufactured byMalvern Instruments, (Worcestershire United Kingdom). Diameters ofparticles reported herein were determining using the Multimodal AnalysisMode of the Zetasizer 1000 to determine particle size by peakintensities. Other techniques may be used to analyze particle size,which results can be correlated to the numerical values obtained withthe light scattering technique described herein.

Addition of the desired passenger molecule occurs prior to dilution withthe solvent if the passenger molecule is lipophilic or amphipathic.Addition occurs after dilution if the passenger molecule is watersoluble.

Thus, in the case of a lipophilic or amphipathic passenger molecule, theNLP loaded populations form upon dilution with the solvent. NLP assemblypopulations or ECVs are formed by dilution of the NLP loaded populationinto water.

In the case of a water soluble passenger molecule, the precursorsolution is mixed with a passenger molecule dissolved in water. NLPassembly populations or ECVs are formed upon dilution with thenon-aqueous solvent. If a serial dilution technique is used, distinctpopulations are formed.

Based on curves observed from different classes of compounds, ranges forthe finished NLP assembly population can be established for each NLPpopulation used to form the final NLP assembly population. The morenon-aqueous solvent that is used to dilute the NLPs, the smaller the NLPassembly populations.

Various NLP loaded populations may be mixed and matched to provide amultifunctional NLP assembly product. The different NLP loadedpopulations within the NLP assembly could provide a preparation whichallows one active ingredient to be preferentially absorbed over theother, thus allowing a control of the rates of release of differentingredients in a single preparation. Alternatively, a single NLPpopulation could be loaded with more than one passenger molecule toprovide the multifunctionality.

Another advantage to the NLP technology is that an optically clearsolution containing NLPs loaded with passenger molecules can be made byselecting conditions where the NLPs are less than about 150 nm in size.It is many times important that a product appear optically clear or itwill fail to gain consumer acceptance. For example, loaded NLPs in anoptically clear solution have application in the beverage industry andthe pharmaceutical industry for liquid products. As one example, amouthwash can be prepared that contains NLPs which encapsulates aningredient for time-release in the mouth. A consumer prefers to purchasean optically clear mouthwash rather than a cloudy one.

The passenger molecules suitable for use in forming a NLP loadedpopulation are numerous. In one embodiment, passenger molecules can beselected which exhibit lipid solubility or are amphipathic. Thesemolecules have solubility profiles ideally suited for loading into NLPS.In another embodiment, water soluble molecules may be incorporated intoNLPs by solubilization into the aqueous solution used to form thefinished NLP product. Using these two approaches virtually any moleculemay be incorporated as a passenger molecule into NLP products of definedsizes. An innovative use of both approaches may be used to incorporateboth lipid and water soluble compounds into a NLP assembly product byfirst incorporating lipid soluble compounds into NLPs prior to dilutionwith ethanol and second incorporating water soluble molecule(s) into thewater solution used to form the finished NLP product of defined size.

NLPs may also be used in the food and beverage industry. For example,NLPs incorporating caffeine may be used in dietary supplements forappetite suppression. Encapsulation in NLPs has been found to beeffective to mask the taste of the passenger molecule if it is desiredthat tasting of such be bypassed upon ingestion.

Another application in the food and beverage industry is theincorporation of substances into NLPs which will be tasted, rather thanmasked. Flavorings such as peppermint oil and other oils areappropriately incorporated into NLPs. The encapsulation ofoil-containing substances may lead to increased shelf life in that theencapsulated substance is protected from oxidation. In addition, theencapsulation of substances would permit additional options formanufacturers and consumers.

As just one example, a manufacturer of a beverage could prepare andbottle one base flavor. The consumer would then have the option ofadding NLP packets to the beverage to meet the taste preferences of theconsumer or to enrich it with vitamins. A consumer that prefers a strongpeppermint flavoring in a chocolate drink could add NLPs containingpeppermint oil to his or her beverage. Substances that are meant to betasted can also be loosely associated with the exterior of the NLP byproviding such substances in the aqueous phase of the procedure. Forexample, an NLP containing a vitamin that preferably should not betasted can have a pleasant taste on the outside thereof.

If it is desired that the NLPs remain in the mouth so that theircontents can be tasted, a natural carbohydrate or sugar can be linked tothe NLP by merely providing it in the aqueous solution. This will stickto the inside of the mouth for a period of time, and normal mouthchemistry and mastication will release the contents of the NLPs toprovide the desired effect. The NLPs can also be subjected to agitationand shear such as in a blender or heavy industrial equipment at amanufacturing site to provide flavorings to foods and beverages.

If the desired passenger molecule is water soluble, the passengermolecule should first be dissolved in water. The incorporation step, orloading of the passenger molecule into the NLP, is accomplished when theNLP product is formed by adding the dissolved passenger molecule to theprecursor solution.

The nanolipidic particles with encapsulated ethanol of the inventionhave a softer “mouth feel” than a preparation containing free ethanol.The encapsulation process leads to the ethanol being sequestered insidethe nanolipid such that the ethanol does not immediately contact themucosa in the mouth. Other passenger molecules which may in thepreparation, such as vitamins and pharmaceutical substances, aresimilarly sequestered within the nanolipidic particles.

Sample Preparation of NLPs and NLP Assembly Populations withEncapsulated Ethanol-Containing Substances

NLPs encapsulating ethanol-containing substances were prepared asfollows:

Solvent-diluted precursor stock was prepared by adding 1 partshelf-stable precursor stock to 0.3 part ethanol to form asolvent-diluted precursor.

An ethanol-containing substance having an ethanol content of 0.2%-50% byvolume is dissolved in an aqueous solvent to form an aqueous-ethanolmonophase. Ethanol-containing substances suitable for encapsulationinclude vodka, gin, rum, bourbon, grain alcohols, or liqueurs containingvodka, gin, rum, bourbon, or grain alcohols.

An aliquot of solvent-diluted precursor stock added to an aliquot of theaqueous-ethanol monophase. This solution is stirred at room temperatureresulting in a loaded NLP population with the desired ethanol-containingsubstance encapsulated within the nanolipid particles to yield aliposomal concentrate comprising ethanol in the amount of about 0.1% to15.0% by volume.

The size of the loaded NLPs may be determined by using the Malvern 1000Zetasizer Laser Light Scattering Instrument set to analyze populationsusing multimodal analysis mode. The size of the finished preparation wasdetermined to be 20 nm-150 nm.

Nanolipid particle sizes useful for the preparation of the invention canbe increased or decreased by adjusting the ratio of ethanol to SolventDilution Microcarrier (SDMC) used in preparation of the precursor stocksolution. Particle sizes can range from approximately 60 nm using 20parts ethanol: 1 part SDMC up to 170 nm using 0.3 part ethanol: 1 partSDMC. Sizes of NLP and NLP assembly populations useful for the method ofthe invention are 20 nm to 300 nm, preferably 20 nm to 170 nm.

One or more additional dilutions of the precursor solution may be madewith ethanol solvent in order to provide a desired size of NLPs andnumber of NLPs per unit volume. The more ethanol solvent that is used todilute the NLPs, the smaller the resulting NLP assembly populations willbe.

In one embodiment, nanolipid particles having ethanol encapsulated at aconcentration of 5.0%-8.0% by volume, is added to base ingredients forgelato. This gelato mixture is then frozen by a commercially acceptableprocess to produce a frozen gelato for consumption.

EXAMPLE 1

Frozen alcohol-containing gelatos having a final ethanol concentrationup to 0.2%-15.0% by volume which have been prepared by the claimedmethod of the invention include the following:

Gelato Flavor % Alcohol by volume Raspberry Cream 8.0% Orange Cream(Grand Marnier ™) 8.0% Bourbon Vanilla 5.0%

In another embodiment, nanolipidic particles having ethanol encapsulatedat a concentration of 5.0% by volume are added to base ingredients for asorbet or frozen beverage. This sorbet or frozen beverage mixture isthen frozen by a commercially acceptable process to produce a frozensorbet, pops, or beverage for consumption. Ice pops and other frozenproducts of a similar nature can be prepared by the same method.

EXAMPLE 2

Frozen alcohol-containing sorbets, pops, and beverages having a finalethanol concentration up to 0.2%-15.0% by volume can be been prepared bythe claimed method of the invention include the following:

Sorbet or Beverage Flavor % Alcohol by volume Piña Colada 5.0% Mojito5.0% Strawberry Margarita 5.0% Apple Martini 5.0%

In yet another embodiment, nanolipid particles having ethanolencapsulated at a concentration of up to 15.0% by volume is added tobase ingredients for a syrup or topping for a frozen dessert.

EXAMPLE 3

Alcohol-containing syrups and toppings having a final ethanolconcentration up to 0.2%-15.0% by volume for frozen desserts which havebeen prepared by the claimed method of the invention include thefollowing:

Syrup or Topping Flavor % Alcohol by volume Caffe (Kahlua ®) 15.0%Chocolate Mint 15.0% Caramel 15.0% Raspberry 15.0% Grand Marnier ® 15.0%Limoncello 15.0%

Stability of NLPs in Commercially Available Alcoholic Beverage Products

NLPs (1:10 Precursor to Ethanol, volume/volume) were prepared anddiluted 1:10 (NLP volume/volume) in commercially available alcoholicbeverage products and stored for one week at Room Temperature. After oneweek the mixtures were vortexed, diluted 1:10 (volume/volume) indistilled water, and the size of the NLPs were analyzed using aZetasizer 1000 (Malvern Instruments). The results of the stability studywere as follows:

Citron ® Vodka 1:10 NLP 150 nm Malibu ® Coconut Rum 1:10 NLP 130 nmBeefeater ® Gin 1:10 NLP 150 nm

Stability of NLPs in 100 Proof (50% volume/volume in Distilled Water)Ethanol Mixtures

NLPs (1:10 and 1:20 Precursor to Ethanol, volume/volume) were preparedand diluted 1:10 in 100 proof mixtures of ethanol and water (50%ethanol, volume/volume). The samples were placed in a commercial freezerfor 14 days, removed, allowed to thaw and warm to Room Temperature. Bothsamples were homogenous and optically clear, without any precipitation.The samples were vortexed and the size of the NLPs were determined usinga Zetasizer 1000 (Malvern Instruments). The results of the analyseswere:

100 Proof Ethanol in Distilled Water Containing NLPs

1:10 NLP 163 nm 1:20 NLP 142 nm

Stability of NLPs After Repeated Freeze Thaw Stored in 25 Proof Ethanolin Distilled Water

NLPs (1:5, 1:10 and 1:20 Precursor to Ethanol volume/volume) wereprepared and added 1:10 (volume/volume) into solutions of 25 ProofEthanol in Distilled Water (12.5% Ethanol in Distilled Water,volume/volume). All mixtures were optically clear. The initial size ofthe NLPs and subsequent size analyses conducted On days 7, 14 and 21were performed using a Zetasizer 1000 (Malvern Instruments). After theinitial size determinations the samples were placed into a commercialfreezer for intervals of 7 days. On days 7, 14 and 21 the samples wereremoved from the freezer allowed to thaw and warm to Room Temperature.

They were vortexed and subjected to size analyses after which they werereturned to the commercial freezer. At days 7, 14 and 21 allpreparations after equilibrating to Room Temperature were opticallyclear and free of any precipitation. The results of the size analyseswere:

NLP Time 0 7 Days 14 Days 21 Days 1:5  156 nm 160 nm 167 nm 162 nm 1:10 77 nm  82 nm  92 nm  94 nm 1:20 105 nm 100 nm 106 nm 116 nm

The NLPs and NLP assembly populations can also be used to formulate adelivery vehicle for pharmaceuticals, such as analgesics, as an admixedpassenger with the NLPs with encapsulated ethanol. The admixed passengerloaded NLPs can then be mixed with ingredients suitable for making afrozen food product. The loaded NLP-frozen food ingredient mixture canbe frozen in a form such as an ice pop to provide a delivery vehicle forthe encapsulated ingredients. One practical application of such adelivery device would be in the treatment of sore throats inindividuals.

The invention can be characterized as follows: a method for makingnanolipid particles (NLPs) having ethanol encapsulated within saidnanolipidic particles, comprising the steps of: a) providing a precursorsolution; b) diluting said precursor solution with an ethanol solvent toproduce a solvent-diluted precursor solution; c) adding anethanol-containing substance having an ethanol content of 0.2%-50.0% byvolume to an aqueous solvent to produce an aqueous-ethanol monophase;and c) mixing said solvent-diluted precursor solution with saidaqueous-ethanol monophase wherein said mixing produces one or morepopulations of ethanol-loaded NLPs or NLP assemblies.

The above invention can also be supplemented as follows: (1) whereinsaid precursor solution is a monophasic optically-clear solution; (2)wherein, said diluting of said precursor solution with said ethanolsolvent is at a ratio ranging from about 1 part precursor to about 20parts solvent to a ratio ranging from about 1 part precursor to about0.3 parts solvent; (3) wherein said NLP assembly has a population formedhaving a mean particle diameter from about 20 nm-300 nm; (4) wherein theconcentration of said solvent in said NLPs is from about 0.5% to about14% by volume; (5) wherein the said concentration of ethanolencapsulated within the nanolipidic particles is from about 0.1% to15.0% by volume; (6) wherein said diluting of said precursor solutionwith said ethanol solvent is at a ratio ranging from about 1 partprecursor to about 20 parts solvent to a ratio ranging from about 1 partprecursor to about 0.3 parts solvent; and; wherein said NLP assemblypopulation is formed having a mean particle diameter from about 20-300nm; and wherein the concentration of said solvent in said NLPs is fromabout 0.5% to about 14% by volume, and wherein the said concentration ofethanol encapsulated within the nanolipidic particles is from about 0.1to 15.0% by volume; (7) wherein said ethanol-containing substance isselected from the group comprising vodka, gin, rum, bourbon, grainalcohol, and liqueurs containing vodka, gin, rum, bourbon, or grainalcohol; (8) wherein the concentration of ethanol in a frozen foodproduct containing ethanol-loaded NLPs is from about 0.1% to 15.0% byvolume; (9) wherein said nanolipidic particles with said encapsulatedethanol-containing substance is combined with ingredients suitable forconsumption in a frozen food product, and the mixture is frozen at anappropriate temperature such that an ethanol-containing frozen foodresults; (10) wherein said ethanol-containing frozen food remains in thefrozen state for a period of time sufficient for an individual toconsume said frozen food; (11) wherein said diluting of said precursorsolution with said ethanol solvent is at a ratio of about 1 partprecursor to about 10 parts solvent to about 1 part loaded nanolipidicpopulation to about 0.5 parts solvent; (12) wherein the stability ofsaid ethanol-encapsulated nanolipidic particles is such that saidparticles will maintain structural integrity through multiplefreeze-thaw cycles; (13) wherein said precursor solution comprisesphospholipids selected from the group consisting of phosphatidylcholine(PC), phosphatidylethanolamine (PE), phosphatidic acid (PA) andphosphatidylinositol (PI) and mixtures thereof; (14) comprising makingone or more additional serial dilutions of said precursor solution withsaid ethanol solvent, wherein said additional dilutions form distinctpopulations of nanolipidic particles, and wherein said nanolipidicparticle populations decrease in size as ethanol concentration inprecursor solution increases; (15) comprising making one or moreadditional dilutions of said precursor solution with said ethanolsolvent in order to provide a desired number of nanolipidic particlesper unit volume; (16) wherein said aqueous solvent further comprises anadditional water-soluble passenger molecule; (17) wherein one or morelipophilic or amphipathic passenger molecules are added to saidprecursor solution to form a loaded nanolipidic particle population,wherein said nanolipidic particles encapsulate admixed passengermolecules; (18) wherein said frozen food is a frozen beverage or desserthaving a final ethanol concentration up to 0.2%-15.0% by volume; (19)wherein said nanolipidic particles with said encapsulatedethanol-containing substance is in a topping for a frozen dessert, saidtopping having a final ethanol concentration of 0.2%-15.0% by volume;and (20) wherein said nanolipidic particles with encapsulated ethanolare admixed with nanolipidic particles encapsulating a pharmaceuticalproduct, said admixture is added to ingredients suitable for a frozenfood, wherein said frozen mixture is a delivery device for saidpharmaceutical.

The invention can alternatively be characterized as follows: acomposition with nanolipidic particles having alcohol-encapsulatedwithin, comprising: a precursor solution, an ethanol solvent thatdilutes said precursor solution and forms a solvent-diluted precursorsolution; an ethanol-containing substance having an ethanolconcentration of 0.2%-50.0% by volume; an aqueous solvent added to theethanol-containing substance to produce an aqueous-ethanol monophase;alcohol encapsulated nanolipid particles formed by the mixture of saidsolvent-diluted precursor solution with said aqueous-ethanol monophase.

This characterization can be supplemented as follows: (1) wherein saidprecursor solution is a monophasic optically-clear solution; (2) whereinsaid precursor solution with said ethanol solvent is at a ratio rangingfrom about 1 part precursor to about 20 parts solvent to a ratio rangingfrom about 1 part precursor to about 0.3 parts solvent: (3) wherein saidNLP assembly has a population formed having a mean particle diameterfrom about 20 nm-300 nm; (4) wherein the concentration of said solventin said NLPs is from about 0.5% to about 14% by volume; (5) wherein thesaid concentration of ethanol encapsulated within the nanolipidicparticles is from about 0.2% to 15.0% by volume; (6) wherein saidprecursor solution with said ethanol solvent is at a ratio ranging fromabout 1 part precursor to about 20 parts solvent to a ratio ranging fromabout 1 part precursor to about 0.3 parts solvent; and wherein said NLPassembly population is formed having a mean particle diameter from about20 nm-300 nm; and wherein the concentration of said solvent in said NLPsis from about 0.5% to about 14% by volume, and wherein the saidconcentration of ethanol encapsulated within the nanolipidic particlesis from about 0.2% to 15.0% by volume; (7) wherein saidethanol-containing substance is selected from the group comprisingvodka, gin, rum, bourbon, grain alcohols, and liqueurs containing vodka,gin, rum, bourbon, or grain alcohols; (8) wherein the concentration ofethanol-containing substance in a frozen food product is from about 0.2%to 15.0% by volume; (9) wherein said nanolipidic particles with saidencapsulated ethanol-containing substance are combined with ingredientssuitable for consumption in a frozen food product, and the mixture isfrozen at an appropriate temperature such that an ethanol-containingfrozen food results; (10) wherein said ethanol-containing frozen foodremains in the frozen state for a period of time sufficient for anindividual to consume said frozen food; (11) wherein said precursorsolution with said ethanol solvent is at a ratio of about 1 partprecursor to about 10 parts solvent to about 1 part loaded nanolipidicpopulation to about 0.5 parts solvent; (12) wherein the stability ofsaid ethanol-encapsulated nanolipidic particles that said particles willmaintain structural integrity through multiple freeze-thaw cycles; (13)wherein said precursor solution comprises phospholipids selected fromthe group consisting of phosphatidylcholine (PC),phosphatidylethanolamine (PE), phosphatidic acid (PA) andphosphatidylinositol (PI) and mixtures thereof; (14) further comprisingone or more additional serial dilutions of said precursor solution,wherein said additional dilutions form distinct populations ofnanolipidic particles, and wherein said nanolipidic particle populationsdecrease in size as ethanol concentration in precursor solutionincreases; (15) further comprising one or more additional dilutions ofsaid precursor solution with said ethanol solvent in order to provide adesired number of nanolipidic particles per unit volume; (16) whereinsaid aqueous solvent has an additional water-soluble passenger molecule;(17) further comprising one or more lipophilic or amphipathic passengermolecules added to said precursor solution to form a loaded nanolipidicparticle population wherein said nanolipidic particles encapsulateadmixed passenger molecules; (18) wherein said nanolipid particles areadded to a frozen beverage or dessert having a final ethanolconcentration from 0.1%-15.0% by volume; (19) wherein said nanolipidparticles are added to a food product, said food product having a finalethanol concentration of from 0.1-15.0% by volume; (20) wherein theethanol encapsulated in said nanolipidic particles is in a concentrationof 0.1% to 15.0%; (21) wherein said nanolipidic particles withencapsulated ethanol are admixed with nanolipidic particlesencapsulating a pharmaceutical product, said admixture is added toingredients suitable for a frozen food, wherein said frozen mixture is adelivery device for said pharmaceutical.

A third characterization of the present invention is a follows: ananolipidic particle (NLP) having encapsulated ethanol, made by aprocess comprising the steps of: a) providing a precursor solution; b)diluting said precursor solution with an ethanol solvent to produce asolvent-diluted precursor solution; c) adding an ethanol-containingsubstance having an ethanol content of 0.2%-50.0% by volume to anaqueous solvent to produce an aqueous-ethanol monophase; and c) mixingsaid solvent-diluted precursor solution with said aqueous-ethanolmonophase wherein said mixing produces one or more populations ofethanol-loaded nanolipids (NLPs) or NLP assemblies.

The third characterization can be supplemented as follows: (1) whereinsaid precursor solution is a monophasic optically-clear solution; (2)wherein said diluting of said precursor solution with said ethanolsolvent is at a ratio ranging from about 1 part precursor to about 20parts solvent to a ratio ranging from about 1 part precursor to about0.3 parts solvent; (3) wherein said NLP assembly has a population formedhaving a mean particle diameter from about 20 nm-300 nm; (4) wherein theconcentration of said solvent in said NLPs is from about 0.5% to about14% by volume; (5) wherein the said concentration of ethanolencapsulated within the nanolipidic particles is from about 0.1% to15.0% by volume; (6) wherein said diluting of said precursor solutionwith said ethanol solvent is at a ratio ranging from about 1 partprecursor to about 20 parts solvent to a ratio ranging from about 1 partprecursor to about 0.3 parts solvent; and; wherein said NLP assemblypopulation is formed having a mean particle diameter from about 20nm-300 nm; and wherein the concentration of said solvent in said NLPs isfrom about 0.5% to about 14% by volume, and wherein the saidconcentration of ethanol encapsulated within the nanolipidic particlesis from about 0.1% to 15.0% by volume; (7) wherein saidethanol-containing substance is selected from the group comprisingvodka, gin, rum, bourbon, grain alcohol, and liqueurs containing vodka,gin, rum, bourbon, or grain alcohol; (8) wherein the concentration ofethanol in said ethanol-loaded NLPs is from about 0.1% to 15.0% byvolume; (9) wherein said nanolipidic particles encapsulated with saidethanol-containing substance are combined with ingredients suitable forconsumption in a frozen food product, and the mixture is frozen at anappropriate temperature such that an ethanol-containing frozen foodresults; (10) wherein said ethanol-containing frozen food remains in thefrozen state for a period of time sufficient for an individual toconsume said frozen food; (11) wherein said diluting of said precursorsolution with said ethanol solvent is at a ratio of about 1 partprecursor to about 10 parts solvent to about 1 part loaded nanolipidicpopulation to about 0.5 parts solvent; (12) wherein the stability ofsaid ethanol-loaded nanolipids is such that said ethanol-loadednanolipids will maintain structural integrity through multiplefreeze-thaw cycles; (13) wherein said precursor solution comprisesphospholipids selected from the group consisting of phosphatidylcholine(PC), phosphatidylethanolamine (PE), phosphatidic acid (PA) andphosphatidylinositol (PI) and mixtures thereof; (14) comprising makingone or more additional serial dilutions of said precursor solution withsaid ethanol solvent, wherein said additional dilutions form distinctpopulations of nanolipidic particles, and wherein said nanolipidicparticle populations decrease in size as ethanol concentration inprecursor solution increases; (15) comprising making one or moreadditional dilutions of said precursor solution with said ethanolsolvent in order to provide a desired number of nanolipidic particlesper unit volume; (16) wherein said aqueous solvent further comprises anadditional water-soluble passenger molecule; (17) wherein one or morelipophilic or amphipathic passenger molecules are added to saidprecursor solution to form a loaded nanolipidic particle populationwherein said nanolipidic particles encapsulate admixed passengermolecules; (18) wherein said frozen food is a frozen beverage or desserthaving a final ethanol concentration of 0.1% to 15.0% by volume; (19)wherein said nanolipidic particles with encapsulated ethanol are in atopping for a frozen dessert, said topping having a final ethanolconcentration of up to of 0.1% to 15.0% by volume; (20) wherein saidnanolipidic particles with encapsulated ethanol are admixed withnanolipidic particles encapsulating a pharmaceutical product, saidadmixture is added to ingredients suitable for a frozen food, whereinsaid frozen mixture is a delivery device for said pharmaceutical.

The examples of ethanol encapsulation in NLPs and NLP assembliespresented herein are representative examples only. The method of theinvention is applicable to other types of ethanol containing substancesand these examples are not meant to constitute the entire range ofethanol-containing substances that may be used in the method disclosedherein.

I claim:
 1. A method for making ethanol-loaded nanolipid particle (NLP)carriers comprising the steps of: a) providing a monophasic opticallyclear precursor stock having phospholipids, ethanol and water; b)diluting said precursor stock with an ethanol solvent to produce asolvent-diluted precursor stock having substantially unloaded nanolipidparticles (NLPs), wherein said diluting of said precursor stock withsaid ethanol solvent is at a ratio ranging from about 1 part precursorstock to about 20 parts ethanol solvent to a ratio ranging from about 1part precursor stock to about 0.3 parts ethanol solvent; c) mixing anethanol-containing substance suitable for human consumption with anaqueous solvent to produce an aqueous-ethanol monophase; and d)combining said solvent-diluted precursor stock produced in step (b) withsaid aqueous-ethanol monophase produced in step (c), wherein theethanol-containing substance from said aqueous-ethanol monophase in step(c) becomes loaded as a passenger within the nanolipid particles (NLPs)in step (b) forming ethanol-loaded nanolipid particle (NLP) carriers. 2.The method of claim 1, wherein said ethanol-containing substancesuitable for human consumption is an ethanol-containing beverageproduct.
 3. The method of claim 1, wherein said ethanol-loaded nanolipidparticle (NLP) carriers are sized from 20 nm-300 nm.
 4. The method ofclaim 1, wherein said aqueous-ethanol monophase in step (c) comprises50% ethanol in water (volume/volume).
 5. The method of claim 1, whereinsaid aqueous-ethanol monophase in step (c) comprises 12.5% ethanol inwater (volume/volume).
 6. The method of claim 1, wherein saidethanol-loaded nanolipidic particle (NLP) carriers are combined with oneor more ingredient suitable for consumption in a food product forming anethanol-containing food product mixture.
 7. The method of claim 5,wherein said ethanol-containing food product mixture is frozen resultingin an ethanol-containing frozen food.
 8. The method of claim 6, whereinsaid ethanol-containing frozen food is a frozen dessert or a frozenbeverage.
 9. The method of claim 5, wherein said ethanol-containing foodproduct mixture is a topping for a frozen food product.
 10. The methodof claim 1, wherein said precursor stock comprises phospholipidsselected from the group consisting of phosphatidylcholine (PC),phosphatidylethanolamine (PE), phosphatidic acid (PA),phosphatidylinositol (PI), and mixtures thereof.
 11. A compositionhaving ethanol-loaded nanolipidic particle (NLPs) carriers comprising: amonophasic optically-clear precursor stock of phospholipids, ethanol andwater; an ethanol solvent added to said precursor stock at a ratio ofabout 1 part precursor stock to about 20 parts ethanol solvent(volume/volume) to about 1 part precursor stock to about 0.3 partsethanol solvent (volume/volume) to result in solvent-diluted precursorstock having one or more substantially unloaded nanolipid particle(NLP); and an aqueous-ethanol monophase made from an ethanol-containingsubstance suitable for human consumption and an aqueous solvent, whereinsaid ethanol-containing substance in said aqueous-ethanol monophaseloads as a passenger within one or more nanolipid particle (NLP) whensaid solvent-diluted precursor stock is combined with saidaqueous-ethanol monophase which results in said ethanol-loaded nanolipidparticle (NLP) carriers.
 12. The composition of claim 11, wherein saidethanol-loaded nanolipid particle (NLP) carriers are sized from 20 nm to300 nm.
 13. The composition of claim 11, wherein said ethanol-containingsubstance suitable for human consumption is an ethanol-containingbeverage product.
 14. The composition of claim 11, wherein saidaqueous-ethanol monophase comprises 50% ethanol in water(volume/volume).
 15. The composition of claim 11, wherein saidaqueous-ethanol monophase comprises 12.5% ethanol in water(volume/volume).
 16. The composition of claim 11, wherein saidethanol-loaded nanolipid particle carriers are combined with ingredientsfor consumption in a food product.
 17. The composition of claim 16,wherein said ingredients are suitable for a frozen food product.
 18. Thecomposition of claim 11, wherein said precursor stock comprisesphospholipids selected from the group consisting of phosphatidylcholine(PC), phosphatidylethanolamine (PE), phosphatidic acid (PA),phosphatidylinositol (PI), and mixtures thereof.
 19. An ethanol-loadednanolipid particle (NLP) carrier made by a process comprising the stepsof: a) providing a monophasic optically clear precursor stock havingphospholipids, ethanol and water; b) diluting said precursor stock withan ethanol solvent to produce a solvent-diluted precursor stock, whereinsaid diluting of said precursor stock with said ethanol solvent is at aratio ranging from about 1 part precursor stock to about 20 partsethanol solvent (volume/volume) to a ratio ranging from about 1 partprecursor stock to about 0.3 parts ethanol solvent (volume/volume), saiddiluting of said precursor stock resulting in one or more substantiallyunloaded nanolipid particle (NLP); c) adding an ethanol-containingbeverage substance for human consumption to an aqueous solvent toproduce an aqueous-ethanol monophase; and d) combining saidsolvent-diluted precursor stock formed in step (b) with saidaqueous-ethanol monophase from step (c), wherein said one or moresubstantially unloaded nanolipid particle (NLP) formed in step (b)becomes loaded with the ethanol-containing substance from step (c) as apassenger in the nanolipid particle (NLP) resulting in one or moreethanol-loaded NLP carrier sized from 20 nm to 300 nm.
 20. The nanolipidparticle (NLP) carrier of claim 19, wherein said ethanol-containingsubstance suitable for human consumption is an ethanol-containingbeverage product.
 21. The nanolipid particle (NLP) carrier of claim 19,wherein one or more ingredient suitable for consumption in a foodproduct are combined with said one or more ethanol-loaded NLP carrier.22. The nanolipid particle (NLP) carrier of claim 19, wherein one ormore ingredient suitable for consumption in a frozen food product arecombined with said one or more ethanol-loaded NLP carrier and frozenresulting in an ethanol-containing frozen food.
 23. The nanolipidparticle (NLP) carrier of claim 19, wherein, said aqueous-ethanolmonophase comprises 12.5% ethanol in water (volume/volume).
 24. Thenanolipid particle (NLP) carrier of claim 19, wherein saidaqueous-ethanol monophase comprises 50% ethanol in water(volume/volume).
 25. The nanolipid particle (NLP) carrier of claim 19,wherein said precursor stock comprises phospholipids selected from thegroup consisting of phosphatidylcholine (PC), phosphatidylethanolamine(PE), phosphatidic acid (PA), phosphatidylinositol (PI), and mixturesthereof.